Immunofluorescence With the ibidi Chambers


ibidi provides several solutions that fit your needs for immunofluorescence assays:

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Chambered Coverslips

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Channel Slides

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Chamber Slides, removable

Bottom material

Glass Coverslip
or Polymer Coverslip

Glass Coverslip
or Polymer Coverslip

Standard glass slide

Additional coverslips required?

No

No

Yes (for mounting)

Microscope type

Inverted

Inverted

Inverted & upright

Mounting medium

Non-hardening

Non-hardening

Hardening

Sample storage

Short-term

Short-term

Long-term

Benefits of ibidi µ-Chambers

  1. High-resolution imaging
    The ibidi slides are ideal for widefield fluorescence, confocal imaging, FRAP, FRET, FLIM, and undisturbed phase contrast imaging.
  2. Fast and simple handling
    The all-in-one chambers simplify your immunofluorescence protocol.
  3. Cost-effective experiments
    Only a small number of cells and low reagent volumes are needed.

With their thin coverslip bottom, the ibidi µ-Dishes and µ-Plates are also ideally suitable for immuno-
fluorescence stainings and high-resolution microscopy.

Comparison of Immunocytochemistry Protocols:
Traditional Staining vs. Staining With ibidi Solutions

When using any of the ibidi solutions, the immunofluorescence staining protocol is much shorter than the traditional protocol. There is no need to grow the cells on loose coverslips—the cells can be grown and stained directly in the ibidi slides.

Protocol With Cells on Coverslips
(Traditional method with nail polish mounting)

  • Sterilize coverslips and slides
  • Coat coverslips
  • Place sterile coverslips into 6-well plate
  • Seed cells in large volume
  • Peel off the coverslip
  • Wash
  • Fix – wash – permeabilize – wash – block
  • Incubate in primary antibody – wash – incubate in secondary antibody
  • Wash
  • Mount cells with mounting medium
  • Mount coverslip with nail polish

Protocol With ibidi µ-Slides
(Time-saving method using all-in-one chambers)

  • Sterilize coverslips and slides
  • Coat coverslips
  • Place sterile coverslips into 6-well plate
  • Seed cells in large volume
  • Peel off the coverslip
  • Wash
  • Fix – wash – permeabilize – wash – block
  • Incubate in primary antibody – wash – incubate in secondary antibody
  • Wash
  • Mount cells with mounting medium
  • Mount coverslip with nail polish

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The ibidi Mounting Medium and the ibidi Mounting Medium With DAPI have a very low autofluorescence, prevent photobleaching and allow the sample to be stored for several weeks on the µ-Slide without the need for additional coverslips.

If your experiment requires the long-term storage of immunostained samples, we recommend using the ibidi Chamber Slides, removable.

Read on and see some Experimental Examples for immunofluorescence, or learn more about a typical Workflow of an Immunofluorescence Staining.